BACKGROUND: Several studies have demonstrated the occurrence of secondary tumors as a rare but significant complication of chimeric antigen receptor T (CAR-T) cell therapy, underscoring the need for a detailed investigation. Given the limited variety of secondary tumor types reported to date, a comprehensive characterization of the various secondary tumors arising after CAR-T therapy is essential to understand the associated risks and to define the role of the immune microenvironment in malignant transformation. This study aims to characterize the immune microenvironment of a newly identified secondary tumor post-CAR-T therapy, to clarify its pathogenesis and potential therapeutic targets. METHODS: In this study, the bone marrow (BM) samples were collected by aspiration from the primary and secondary tumors before and after CD19 CAR-T treatment. The CD45 + BM cells were enriched with human CD45 microbeads. The CD45 + cells were then sent for 10× genomics single-cell RNA sequencing (scRNA-seq) to identify cell populations. The Cell Ranger pipeline and CellChat were used for detailed analysis. RESULTS: In this study, a rare type of secondary chronic myelomonocytic leukemia (CMML) were reported in a patient with diffuse large B-cell lymphoma (DLBCL) who had previously received CD19 CAR-T therapy. The scRNA-seq analysis revealed increased inflammatory cytokines, chemokines, and an immunosuppressive state of monocytes/macrophages, which may impair cytotoxic activity in both T and natural killer (NK) cells in secondary CMML before treatment. In contrast, their cytotoxicity was restored in secondary CMML after treatment. CONCLUSIONS This finding delineates a previously unrecognized type of secondary tumor, CMML, after CAR-T therapy and provide a framework for defining the immune microenvironment of secondary tumor occurrence after CAR-T therapy. In addition, the results provide a rationale for targeting macrophages to improve treatment strategies for CMML treatment.
Humans ; Lymphoma, Large B-Cell, Diffuse/therapy* ; Tumor Microenvironment/genetics* ; Antigens, CD19/metabolism* ; Leukemia, Myelomonocytic, Chronic/genetics* ; Immunotherapy, Adoptive/adverse effects* ; Male ; Single-Cell Analysis/methods* ; Female ; Sequence Analysis, RNA/methods* ; Receptors, Chimeric Antigen ; Middle Aged

Objective To compare the efficacy and tolerability of the novel bowel-cleansing agent TCIC-001 and the traditional polyethylene glycol (PEG) regimen for bowel preparation prior to colonoscopy. Methods Prospective inclusion of 62 patients who were scheduled to undergo colonoscopy at Zhongshan Hospital, Fudan University from July 2021 to July 2022. They were randomly divided into TCIC-001 group (n=31) and PEG group (n=31) using a random number table method. The TCIC-001 group took TCIC-001 orally, drinking water in stages, with a total liquid intake of 1 500 mL; the PEG group took PEG orally, taking it in 4 doses, with a total liquid intake of 3 000 mL. The primary endpoint indicator is the quality of intestinal hygiene evaluated by the Boston Bowel Preparation Scale (BBPS), the secondary endpoint indicators were medication adherence, medication duration, frequency of bowel movements, duration of bowel movements, and incidence of adverse events between two groups. Results No significant differences were observed in sex, age, or defecation frequency between the two groups. For efficacy, both groups achieved equivalent bowel cleanliness, with a “good preparation” rate of 93.55% and comparable BBPS score of each intestinal segment and total scores. For tolerability, the TCIC-001 group had a shorter medication duration compared to the PEG group ([48.8±25.9] min vs [82.8±28.4] min, P<0.001), a longer defecation duration ([288.6±74.0] min vs [236.5±74.3] min, P<0.001), and a lower incidence of first defecation before medication completion (9.68% vs 41.94%, P=0.004). Regarding safety, no significant differences were observed between the TCIC-001 group and the PEG group in incidences of chloride disturbances (0% vs 9.68%) and calcium disturbances (3.23% vs 6.45%), and no other adverse events. Conclusions TCIC-001 demonstrated comparable bowel-cleansing efficacy to PEG while significantly improving tolerability (reduced medication time and lower risk of premature defecation) and maintaining favorable safety.

Objective:To elucidate the factors influencing the differences in the survival rates of esophageal squamous cell carcinoma (ESCC) patients between the rural and urban regions in China. Methods:A total of 36,723 ESCC patients derived from the clinical data-bases containing 500,000 esophageal and gastric cardia carcinoma cases (1973-2015) of the Henan Key Laboratory for Esophageal Can-cer Research of the First Affiliated Hospital, Zhengzhou University, were analyzed. Of these patients, 33,625 were from the rural re-gions (91.6%), comprising 20,906 male patients with an average age of 58.98 ± 8.71 years and 12,719 females with an average age of 59.59 ± 8.53 years. The remaining 3,098 were from the urban regions and composed of 2,089 male patients with an average age of 60.84±9.10 years and 1,009 females with an average age of 62.46 ± 9.14 years. All the patients underwent radical esophagectomy, de-tailed histopathological diagnosis, and TNM staging. Chi square test, Kaplan-Meier, Log-rank, and Cox proportional hazards regression model were used to analyze the differences between ESCC patients from rural regions and those from urban regions and among the risk factors in prognosis. Results:Kaplan-Meier and Log-rank analysis results showed that the ESCC patients from the rural regions had significantly higher overall survival than the urban patients (χ2=12.971, P=0.000). Further analysis showed that rural patients≥50 years old and diagnosed with stage IIa and IIb (middle stage) ESCC had higher survival rates than urban patients in males and females (male:χ2=16.188, P<0.001;female:χ2=5.019, P=0.025). However, the survival rates of rural and urban patients with stage 0,Ⅰa,Ⅰb (early stage) and Ⅲa, Ⅲc, and Ⅳ (late stage) were similar (P>0.05). The results of Cox proportional hazards regression model analysis showed that age, gender, and TNM stages were independent risk factors for rural and urban ESCC patients. When the rural and urban ESCC patients were both considered, the Cox proportional hazards regression model analysis results showed that male ESCC patients≥50 years old, urban residence, and TNM stages were independent risk factors. Conclusion:Rural ESCC patients have significantly high-er overall survival than urban patients. Male, age of≥50 years old, urban residence, and TNM stages were independent risk factors for ESCC patient survival.

Objective To investigate the correlation of bone mineral density (BMD) with plasma Klotho levels and its related factors in type 2 diabetic patients (T2DM) . Methods BMD was measured by Dual-energy X-ray absorptiometry (DEXA) in 159 T2DM patients. The patients were divided into three groups:normal bone mass, reduced bone mass and osteoporosis. The fasting plasma levels of Klotho were detected in these patients using enzyme linked immuno sorbent assay (ELISA), clinical and biochemical parameters also were tested, the difference and related factors were compared and analyzed in each group. Results Plasma Klotho levels were not significantly different among the three groups (4.95±0.48 vs 4.96±0.47 vs 4.91±0.49,P>0.05) . BMD at the first, second, third, fourth and total lumbar spine, femoral neck, trochanter and total body were not associated with plasma Klotho levels in these patients (P>0.05) . Age, diabetic duration, HDL-C and BMI were independent determinants for BMD in T2DM patients. Conclusions BMD might be not associated with plasma Klotho level in T2DM patients. But age,diabetic duration,HDL-C and BMI are associated with reduced BMD and osteoporosis in T2DM patients.

Objective To explore the effects of microRNA(miR)-1283 on invasion,proliferation and apoptosis of HTR 8/SVneo cell line derived from human-trophoblast cells.Methods HTR-8/SVneo cells were divided into three groups,as-miR 1283 group was transfected with miR 1283 analogues,anti-miR-1283 group was transfected with miR-1283 inhibitors,and negative control group was transfected with non functional sequence.The levels of miR 1283 expression were detected by fluorescence quantitative polymerase chain reaction at 24 hours after transfection.The cell proliferation was measured by 3-(4,5)-dimethylthiazol-2-yl-(2,5)-diphenyl tetrazolium bromide assay at 24,48 and 72 hours after transfection.Apoptosis was evaluated by propidium iodide staining and flow cytometry at 48 hours after transfection.Transwell experiments were performed to evaluate cellular invasion abilities at 24 hours after transfection.Analysis of variance and Bonferroni method were applied as statistical methods.Results The level of miR 1283 in as miR 1283 group was higher than that in the negative control group with statistically significant difference (14.85±0.57 vs 7.08±0.20,P＜0.01).At 24,48 and 72 hours after transfection,the inhibitory rate of cell proliferation in as-miR-1283 group was (8.04 ± 0.27) ％,(32.47 ± 0.24) ％ and (9.23± 0.17) ％,higher than those in the negative control group [(0.72 ± 0.06) ％,(1.17 ± 0.04) ％ and (0.53 ± 0.05) ％] (P ＜ 0.01,respectively).Cell apoptosis rate was higher in as-miR 1283 group than in negative control group [(16.33 ± 0.40) ％ vs (9.39± 0.58) ％,P＜0.01].The transmembrane cell number was lower in as-miR-1283 group as comparing with the negative control group (7.25 ± 1.83 vs 16.33 ± 2.08,P＜0.01).miR-1283 expression,apoptosis and transmembrane cell number in anti-miR-1283 group had no statistical difference as compared to the negative control group (all P ＞ 0.05).Conclusions Up-regulated levels of miR-1283 could inhibit HTR-8/SVneo cell proliferation and invasion,but promote the cell apoptosis.

Objective To summarize our experience in the management of cardiopulmonary bypass (CPB) with deep hypothermic circulatory arrest (DHCA) during aortic arch surgery.Methods From March 2007 to May 2010,58 consecutive patients,including 24 urgent and 34 selective operations underwent aortic arch surgery.Thirty-nine hemiarch and 19 total aortic arch replacement operations were performed.CPB was established by perfusion through femoral artery (42 cases) and right subclavian artery (RSA) ( 16 cases),of which 4 cases were carried out with antegrade cerebral perfusion (ACP).Results The mean CPB time was ( 208.88 ± 136.45 ) min.The mean cerebral circulation arrest was ( 27.36 ± 11.50 ) min.Nasopharyngeal and rectal temperature were ( 16.01 ±2.67)℃ and ( 19.72 ±2.13)℃ respectively before DHCA was initiated.The mean times for cooling and rewarming were ( 50.91 ± 16.89) min and ( 88.97 ± 43.68 ) min.The mean time of intubation was (56.70 ± 45.19 ) h.The time in ICU was ( 5.68 ± 5.31 ) d,and the time of hospitalization was (30.11 ± 22.27 ) d.Acute renal failure,hypoxemia,and paraplegia occurred post-operatively in 4,19,and 2 patients,respectively.Four patients died post-operatively with a mortality of 6.90％.Compared with those received hemiarch replacement operation,the patients received total aortic arch replacement had statistically longer time of CPB([262.16 ±219.97]min vs [182.92 ±53.81] min,t =2.14,P ＜0.05),cerebral circulatory arrest ( [30.47 ± 15.86 ] win vs [25.85 ± 8.48 ] min,t =2.40,P ＜ 0.05 ),rewarming ( [110.00 ± 68.66 ] min vs [78.72 ± 17.31 ] min,t =2.69,P ＜ 0.05 ),and intubation ( [93.95 ± 131.89 ] h vs [38.08 ± 30.70 ] h,t =2.50,P ＜ 0.05 ).There was no significant difference in the times of these procedures between emergency surgery group and elective surgery group,between RSA and femoral artery cannulation groups.Conclusion It is crucial that the cooling and re-warming procedures during aortic arch surgery should be carried out slowly,gradually,and completely when DHCA was adopted alone.conclusion through right axillary artery or RSA was preferred for ACP,in order to accomplish the body circulation arrest at a relative high temperature,to shorten the CPB time,and to alleviate potential harmful effects of hypothermia.Meticulous management of CPB is one of the most important measures to improve the patients' outcome.

Objective: To study the effects of genistein on the proliferation and cell cycle progression of human gastric carcinoma cells. Methods: 3H TdR incorporation test was used to investigate the cell proliferation. Flow cytometry was used to analyze the cell cycle arrest. Immunocytochemistry technique and Western blotting were used to observe the cyclin B and P21 waf1/cip1 protein expression. Results: Genistein inhibited the proliferation of tumor cells significantly, arrested cell cycle progression at G 2/M phase, and enhanced cyclin B and P21 waf1/cip1 protein expression in dose dependent manner. Conclusion:Proliferatory inhibition and G 2/M arrest of human gastric carcinoma cells after treated with genistein may be due to increased stability of cyclin B protein and the expression of P21 waf1/cip1 .